PURIFICATION AND CHARACTERIZATION OF HUMAN CHITOTRIOSIDASE, A NOVEL MEMBER OF THE CHITINASE FAMILY OF PROTEINS
PURIFICATION AND CHARACTERIZATION OF HUMAN CHITOTRIOSIDASE, A NOVEL MEMBER OF THE CHITINASE FAMILY OF PROTEINS
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DOI：
10.1074/jbc.270.5.2198
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发表时间：
1995-02-03
影响因子：
4.8
通讯作者：
AERTS, JMFG
中科院分区：
文献类型：
--
作者：
RENKEMA, GH;BOOT, RG;AERTS, JMFG
最近我们注意到（Hollak，C. E. M.，货车Weely，S.，货车Oers，M. H. J.，和Awesley，J. M. F. G.等人（1994）J. Clin. Invest. 93，1288-1292），戈谢病的临床表现与血浆中壳三糖苷酶活性的几百倍增加有关，我们报道了该蛋白的纯化和表征。从戈谢病患者的脾中纯化了壳三糖苷酶的两种主要同种型，其等电点分别为7.2和8.0，分子量分别为50和39 kDa，两种形式的N-末端氨基酸序列证明是相同的，抗纯化的39-kDa壳三糖苷酶的抗血清沉淀所有同工酶，壳三糖苷酶活性在一些个体中早期发现完全不存在，这些发现组合表明，单个基因可能编码壳三糖苷酶的不同亚型。末端序列和内部序列壳三糖苷酶被证明与作为几丁质酶家族成员的蛋白质中的序列同源（Hakala，B. E、白色，C.和Recklies，A. D.（1993）J.Biol.Chem.268，25803-25810），本文所述的人壳三糖苷酶显示出对人工底物以及几丁质的几丁质分解活性，因此可以认为是几丁质酶。
Recently we noted (Hollak, C. E. M., van Weely, S., van Oers, M. H. J., and Aerts, J. M. F. G. (1994) J. Clin. Invest. 93, 1288-1292) that the clinical manifestation of Gaucher disease is associated with a several hundred-fold increase in chitotriosidase activity in plasma, We report on the purification and characterization of the protein.Two major isoforms of chitotriosidase with isoelectric points of 7.2 and 8.0 and molecular masses of 50 and 39 kDa, respectively, were purified from the spleen of a Gaucher patient, The N-terminal amino acid sequence of the two forms proved to be identical, An antiserum raised against the purified 39-kDa chitotriosidase precipitated all isozymes, Chitotriosidase activity was earlier found to be completely absent in some individuals, These findings in combination suggest that a single gene may encode the different isoforms of chitotriosidase.Both the N-terminal sequence and an internal sequence chitotriosidase proved to be homologous to sequences in proteins that are members of the chitinase family (Hakala, B. E., White, C., and Recklies, A. D. (1993) J. Biol. Chem. 268, 25803-25810), The human chitotriosidase described here showed chitinolytic activity toward artificial substrates as well as chitin and may therefore be considered to be a chitinase.